Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.
Explain the difference in results for the RT-PCR from the CD4+ T cells and the RT-PCR from the CD8+
T cells. (i.e., propose a conclusion)
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The structure indicates the following: the start and end of transcription, the start codon and the stop
codon, the location of the two introns (marked with black boxes). The white boxes indicate the binding
locations of two PCR primers.
PCR CD4+ T cells
transcriptional
start site
RT-PCR CD4+ T cells
+1
RT-PCR CD8+ T cells
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Size markers
The researchers carry out a FACS (Fluorescence Activated Cell Sorting) separation of human blood which
isolates a relatively pure sample of CD4+ T cells, and a relatively pure sample of CD8+ T cells, both
different types of lymphocytes. They carry out a PCR, and a semi-quantitative RT-PCR, on these cell
samples. The results are shown on the gel.
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intron
DNA migration
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intron
transcriptional
stop site
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